Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
PERK Double Nickase Plasmid (h): sc-400080-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PERK Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- PERK Double Nickase Plasmid (h) and PERK Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EIF2AK3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PERK Antibody (B-5): sc-377400
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PERK Double Nickase Plasmid (h) | sc-400080-NIC | 20 µg | $410.00 | |||
PERK Double Nickase Plasmid (h2) | sc-400080-NIC-2 | 20 µg | $410.00 |
EIF2AK3 encodes PERK, an ER-resident transmembrane kinase that senses unfolded protein accumulation and initiates the unfolded protein response. Upon activation, PERK phosphorylates eIF2α to attenuate global translation while promoting selective translation programs, thereby coordinating proteostasis, oxidative stress control, and autophagy-related adaptations. PERK signaling intersects with ATF4/CHOP-mediated transcriptional outputs and influences ER–mitochondrial crosstalk and redox homeostasis. Dysregulated PERK activity has been associated with chronic ER stress states implicated in neurodegeneration, metabolic dysfunction, and tumor cell stress adaptation, making it a key node for mechanistic studies of stress signaling.
PERK Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EIF2AK3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EIF2AK3. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt EIF2AK3 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of EIF2AK3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.